Frances Carol Arnold

US Patent:

6361988, Mar 26, 2002

Filed:

Apr 4, 2000

Appl. No.:

09/543084

Inventors:

Frances H. Arnold - Pasadena CA
Zhixin Shao - Penzberg, DE
Huimin Zhao - San Diego CA
Lorraine J. Giver - Sunnyvale CA

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

C12N 916

US Classification:

435196, 4352523, 4353201, 435 912, 435 915, 536 232

Abstract:

A method for in vitro mutagenesis and recombination of polynucleotide sequences based on polymerase-catalyzed extension of primer oligonucleotides is disclosed. The method involves priming template polynucleotide(s) with random-sequences or defined-sequence primers to generate a pool of short DNA fragments with a low level of point mutations. The DNA fragments are subjected to denaturization followed by annealing and further enzyme-catalyzed DNA polymerization. This procedure is repeated a sufficient number of times to produce full-length genes which comprise mutants of the original template polynucleotides. These genes can be further amplified by the polymerase chain reaction and cloned into a vector for expression of the encoded proteins.

US Patent:

6524837, Feb 25, 2003

Filed:

Feb 3, 2000

Appl. No.:

09/497585

Inventors:

Frances H. Arnold - Pasadena CA
Oliver May - Stuttgart, DE
Karlheinz Drauz - Freigericht, DE
Andreas Bommarius - Frankfurt/M, DE

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

C12N 986

US Classification:

435231

Abstract:

Hydantoinase enzymes which are mutants of a previously isolated hydantoinase having the amino acid SEQ. ID. NO. 2. The mutants include amino acid substitutions at positions 95, 154, 180, 251 and/or 255 of the wild type hydantoinase (SEQ. ID. NO. 2). The mutant hydantoinases, like the parent hydantoinase, are used in the production of optically pure amino acids.

US Patent:

6537746, Mar 25, 2003

Filed:

Dec 4, 1998

Appl. No.:

09/205448

Inventors:

Frances Arnold - Pasadena CA
Zhixin Shao - Penzberg DE
Alexander Volkov - South Pasadena CA

Assignee:

Maxygen, Inc. - Redwood City CA

International Classification:

C12Q 168

US Classification:

435 6, 435 911, 435 912, 435 691, 4353201, 536 27, 935 47

Abstract:

The invention provides methods for evolving a polynucleotide toward acquisition of a desired property. Such methods entail incubating a population of parental polynucleotide variants under conditions to generate annealed polynucleotides comprising heteroduplexes. The heteroduplexes are then exposed to a cellular DNA repair system to convert the heteroduplexes to parental polynucleotide variants or recombined polynucleotide variants. The resulting polynucleotides are then screened or selected for the desired property.

US Patent:

6540895, Apr 1, 2003

Filed:

May 21, 1999

Appl. No.:

09/325667

Inventors:

Charles F. Spence - Arcadia CA
Anne Y. Fu - Hacienda Heights CA
Stephen R. Quake - San Marino CA
Frances H. Arnold - Pasadena CA

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

G01N 2726

US Classification:

204450, 204451, 204600, 204601, 4352831, 4352861, 4352865, 4352866

Abstract:

The invention provides a microfabricated device for sorting cells based on a desired characteristic, for example, reporter-labeled cells can be sorted by the presence or level of reporter on the cells. The device includes a chip having a substrate into which is microfabricated at least one analysis unit. Each analysis unit includes a main channel, having a sample inlet channel, typically at one end, and a detection region along a portion of its length. Adjacent and downstream from the detection region, the main channel has a discrimination region or branch point leading to at least two branch channels. The analysis unit may further include additional inlet channels, detection points, branch points, and branch channels as desired. A stream containing cells is passed through the detection region, such that on average one cell occupies the detection region at a given time. The cells can be sorted into an appropriate branch channel based on the presence or amount of a detectable signal such as an optical signal, with or without stimulation, such as exposure to light in order to promote fluorescence.

US Patent:

6902918, Jun 7, 2005

Filed:

Feb 9, 1999

Appl. No.:

09/246451

Inventors:

Frances H. Arnold - Pasadena CA, US
Hyun Joo - Suwon, KR
Zhanglin Lin - Beijing, CN

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

C12N009/00
C12N009/02
C12N001/20
C12N015/00
C07H021/04

US Classification:

435189, 435 4, 435 6, 435 25, 435 691, 435183, 4352523, 4353201, 536 232, 536 234, 536 237

Abstract:

A method for detecting the presence of an oxygenated compound which is produced when a substrate is reacted with an oxygenase for the substrate. The method involves reacting a coupling enzyme with the oxygenated compound to form a polymeric oxygenated compound which is fluorescent or luminescent. Measurement of the fluorescence or luminescence of the polymeric oxygenated compound provides indirect detection of the oxygenated compound produced by reaction of the oxygenase with the substrate. The method is carried out in a whole cell environment wherein the cell is transformed to express both the oxygen a set being screened and the coupling enzyme. The method can be used to measure the activity of monooxygenases and dioxygenases on aromatic substrates. The method is amenable to large scale screening of enzyme mutants to isolate those with maximum oxygenase activity.

US Patent:

7098010, Aug 29, 2006

Filed:

Nov 27, 2000

Appl. No.:

09/722602

Inventors:

Frances H. Arnold - Pasadena CA, US
Ioanna P. Petrounia - Pasadena CA, US
Lianhong Sun - Pasadena CA, US

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

C12N 9/02
C12N 1/20
C12N 15/00
C12Q 1/68
C12Q 1/00
C12P 21/04
C07H 21/04

US Classification:

435189, 435 4, 435 6, 435 691, 435 711, 435440, 4352523, 4353201, 536 232, 536 237

Abstract:

This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantagoeusly produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.

US Patent:

7115403, Oct 3, 2006

Filed:

May 16, 2000

Appl. No.:

09/571553

Inventors:

Frances H. Arnold - Pasadena CA, US
Ioanna P. Petrounia - Pasadena CA, US
Lianhong Sun - Pasadena CA, US

Assignee:

The California Institute of Technology - Pasadena CA

International Classification:

C12N 9/02
C12N 1/20
C12N 15/00
C12Q 1/68
C12Q 1/00
C07H 21/04
C12P 21/04

US Classification:

435189, 435 4, 435 6, 435 691, 435 711, 435440, 4352523, 4353201, 536 232, 536 237

Abstract:

This invention relates to the expression of improved polynucleotide and polypeptide sequences encoding for eukaryotic enzymes, particularly oxidase enzymes. The enzymes are advantageously produced in conventional or facile expression systems. Various methods for directed evolution of polynucleotide sequences can be used to obtain the improved sequences. The improved characteristics of the polypeptides or proteins generated in this manner include improved expression, enhanced activity toward one or more substrates, and increased thermal stability. In a particular embodiment, the invention relates to improved expression of the galactose oxidase gene and galactose oxidase enzymes. GAO mutants that are highly active and/or thermostable are disclosed.

US Patent:

7166466, Jan 23, 2007

Filed:

Feb 19, 2003

Appl. No.:

10/371168

Inventors:

Frances Arnold - Pasadena CA, US
Zhixin Shao - Penzberg DE, US
Alexander Volkov - South Pasadena CA, US

Assignee:

California Institute of Technology - Pasadena CA

International Classification:

C12N 15/00
C12Q 1/68
C07H 21/02
C07H 21/04

US Classification:

435440, 435 6, 536 231, 536 243

Abstract:

The invention provides methods for evolving a polynucleotide toward acquisition of a desired property. Such methods entail incubating a population of parental polynucleotide variants under conditions to generate annealed polynucleotides comprising heteroduplexes. The heteroduplexes are then exposed to a cellular DNA repair system to convert the heteroduplexes to parental polynucleotide variants or recombined polynucleotide variants. The resulting polynucleotides are then screened or selected for the desired property.